{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T20:04:30Z","timestamp":1782763470602,"version":"3.54.5"},"reference-count":51,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2025,6,24]],"date-time":"2025-06-24T00:00:00Z","timestamp":1750723200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Metabolites"],"abstract":"<jats:p>Background\/Objectives: Non-alcoholic steatohepatitis (NASH) carries a high risk of developing hepatic fibrosis. Hugan tablets (HGTs), a traditional Chinese medicine, have exhibited potent anti-hepatic fibrosis effects, though the underlying mechanisms remain unclarified. This study aims to assess the efficacy of HGTs against NASH-related liver fibrosis in mice and investigate the underlying mechanisms via the integration of pseudotargeted metabolomics and microbiomics. Methods: C57BL\/6 mice were fed a choline-deficient, ethionine-supplemented (CDE) diet and treated with HGTs. The therapeutic effects of HGTs in CDE mice were assessed. The underlying mechanism of HGTs was investigated by the integration of microbiomics, a pseudo-sterile model, untargeted followed by pseudotargeted metabolomics, and molecular docking. Results: HGTs alleviated NASH-related hepatic fibrosis in CDE mice and restored the composition of the gut microbiota. The depletion of the gut microbiota eliminated the anti-hepatic fibrosis effect of HGTs. HGTs increased intestinal 7-ketolithocholic acid and tauroursodeoxycholic acid via 7\u03b1\/\u03b2-hydroxysteroid dehydrogenase (7\u03b1\/\u03b2HSDH), while reducing deoxycholic acid (DCA) and taurodeoxycholic acid through inhibition of bile acid 7\u03b1-dehydratase (BaiE), leading to lower hepatic DCA. Six intestinal components of HGTs interacted with 7\u03b1HSDH, 7\u03b2HSDH, and BaiE, which are expressed in the bacterial genera altered by HGTs. Conclusions: HGTs alleviate NASH fibrosis by reshaping the gut microbiome, acting on microbial BA-metabolizing enzymes, and regulating the BA metabolism in the liver and gut.<\/jats:p>","DOI":"10.3390\/metabo15070433","type":"journal-article","created":{"date-parts":[[2025,6,24]],"date-time":"2025-06-24T09:24:18Z","timestamp":1750757058000},"page":"433","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Integration of Pseudotargeted Metabolomics and Microbiomics Reveals That Hugan Tablets Ameliorate NASH with Liver Fibrosis in Mice by Modulating Bile Acid Metabolism via the Gut Microbiome"],"prefix":"10.3390","volume":"15","author":[{"given":"Wenran","family":"Dong","sequence":"first","affiliation":[{"name":"School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huajinzi","family":"Li","sequence":"additional","affiliation":[{"name":"School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huilin","family":"Ma","sequence":"additional","affiliation":[{"name":"School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingxi","family":"Gong","sequence":"additional","affiliation":[{"name":"School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gan","family":"Luo","sequence":"additional","affiliation":[{"name":"School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyan","family":"Gao","sequence":"additional","affiliation":[{"name":"School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"101231","DOI":"10.1016\/j.mam.2023.101231","article-title":"Liver fibrosis in NAFLD\/NASH: From pathophysiology towards diagnostic and therapeutic strategies","volume":"95","author":"Parola","year":"2024","journal-title":"Mol. Aspects Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1002\/hep.29367","article-title":"The diagnosis and management of nonalcoholic fatty liver disease: Practice guidance from the American Association for the Study of Liver Diseases","volume":"67","author":"Chalasani","year":"2018","journal-title":"Hepatology"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1053\/j.gastro.2023.05.053","article-title":"Prevalence of Liver Steatosis and Fibrosis in the General Population and Various High-Risk Populations: A Nationwide Study With 5.7 Million Adults in China","volume":"165","author":"Man","year":"2023","journal-title":"Gastroenterology"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1154","DOI":"10.1016\/j.cgh.2022.07.033","article-title":"Disease State Transition Probabilities Across the Spectrum of NAFLD: A Systematic Review and Meta-Analysis of Paired Biopsy or Imaging Studies","volume":"21","author":"Le","year":"2023","journal-title":"Clin. Gastroenterol. Hepatol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/S2468-1253(23)00275-3","article-title":"Hepatocellular and extrahepatic cancer risk in people with non-alcoholic fatty liver disease","volume":"9","author":"Thomas","year":"2024","journal-title":"Lancet Gastroenterol. Hepatol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1038\/s41575-023-00807-x","article-title":"Hepatic inflammatory responses in liver fibrosis","volume":"20","author":"Hammerich","year":"2023","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2717","DOI":"10.1016\/j.cell.2024.03.034","article-title":"Gut symbionts alleviate MASH through a secondary bile acid biosynthetic pathway","volume":"187","author":"Nie","year":"2024","journal-title":"Cell"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"348","DOI":"10.1038\/s41575-024-00896-2","article-title":"Another renaissance for bile acid gastrointestinal microbiology","volume":"21","author":"Ridlon","year":"2024","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.mam.2017.04.004","article-title":"The role of bile acids in nonalcoholic fatty liver disease and nonalcoholic steatohepatitis","volume":"56","author":"Chow","year":"2017","journal-title":"Mol. Aspects Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1811","DOI":"10.1002\/hep.32568","article-title":"Metabolic reprogramming of the intestinal microbiome with functional bile acid changes underlie the development of NAFLD","volume":"76","author":"Smirnova","year":"2022","journal-title":"Hepatology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1002\/hep.29359","article-title":"The presence and severity of nonalcoholic steatohepatitis is associated with specific changes in circulating bile acids","volume":"67","author":"Puri","year":"2018","journal-title":"Hepatology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2323233","DOI":"10.1080\/19490976.2024.2323233","article-title":"BaiJ and BaiB are key enzymes in the chenodeoxycholic acid 7\u03b1-dehydroxylation pathway in the gut microbe Clostridium scindens ATCC 35704","volume":"16","author":"Meibom","year":"2024","journal-title":"Gut Microbes"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1038\/nchembio.1864","article-title":"A biosynthetic pathway for a prominent class of microbiota-derived bile acids","volume":"11","author":"Devlin","year":"2015","journal-title":"Nat. Chem. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Guzior, D.V., and Quinn, R.A. (2021). Review: Microbial transformations of human bile acids. Microbiome, 9.","DOI":"10.1186\/s40168-021-01101-1"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Heinken, A., Ravcheev, D.A., Baldini, F., Heirendt, L., Fleming, R.M.T., and Thiele, I. (2019). Systematic assessment of secondary bile acid metabolism in gut microbes reveals distinct metabolic capabilities in inflammatory bowel disease. Microbiome, 7.","DOI":"10.1186\/s40168-019-0689-3"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1038\/s41591-019-0406-6","article-title":"Meta-analysis of fecal metagenomes reveals global microbial signatures that are specific for colorectal cancer","volume":"25","author":"Wirbel","year":"2019","journal-title":"Nat. Med."},{"key":"ref_17","first-page":"2943","article-title":"Expert consensus statement on Hugan Tablets in clinical practice","volume":"44","author":"Liu","year":"2019","journal-title":"China J. Chin. Mater. Medica"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Jiang, Y., Chen, M., Gang, H., Li, X., Zhai, C., Feng, Z., Luo, G., and Gao, X. (2023). A funnel-type stepwise filtering strategy for identification of potential Q-markers of traditional Chinese medicine formulas. Front. Pharmacol., 14.","DOI":"10.3389\/fphar.2023.1143768"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Lv, S., Yu, H., Liu, X., and Gao, X. (2021). The Study on the Mechanism of Hugan Tablets in Treating Drug-Induced Liver Injury Induced by Atorvastatin. Front. Pharmacol., 12.","DOI":"10.3389\/fphar.2021.683707"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1080\/13880209.2018.1479868","article-title":"Effects of garlic polysaccharide on alcoholic liver fibrosis and intestinal microflora in mice","volume":"56","author":"Wang","year":"2018","journal-title":"Pharm. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1038\/s41419-019-1678-y","article-title":"Hepatic ferroptosis plays an important role as the trigger for initiating inflammation in nonalcoholic steatohepatitis","volume":"10","author":"Tsurusaki","year":"2019","journal-title":"Cell Death Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.jhep.2006.08.015","article-title":"Attenuated liver progenitor (oval) cell and fibrogenic responses to the choline deficient, ethionine supplemented diet in the BALB\/c inbred strain of mice","volume":"46","author":"Knight","year":"2007","journal-title":"J. Hepatol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1625","DOI":"10.1002\/hep.22820","article-title":"Relation between liver progenitor cell expansion and extracellular matrix deposition in a CDE-induced murine model of chronic liver injury","volume":"49","author":"Sempoux","year":"2009","journal-title":"Hepatology"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1239","DOI":"10.1136\/gutjnl-2019-319114","article-title":"Myristoleic acid produced by enterococci reduces obesity through brown adipose tissue activation","volume":"69","author":"Quan","year":"2020","journal-title":"Gut"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1111\/bph.15965","article-title":"Aberrant branched-chain amino acid accumulation along the microbiota-gut-brain axis: Crucial targets affecting the occurrence and treatment of ischaemic stroke","volume":"180","author":"Shen","year":"2023","journal-title":"Br. J. Pharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1174","DOI":"10.1136\/gutjnl-2020-323071","article-title":"Gut microbiota-derived metabolites as central regulators in metabolic disorders","volume":"70","author":"Agus","year":"2021","journal-title":"Gut"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"584","DOI":"10.1016\/j.molcel.2020.03.005","article-title":"From Association to Causality: The Role of the Gut Microbiota and Its Functional Products on Host Metabolism","volume":"78","author":"Koh","year":"2020","journal-title":"Mol. Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1038\/s41575-020-0269-9","article-title":"Gut microbiota and human NAFLD: Disentangling microbial signatures from metabolic disorders","volume":"17","author":"Vigliotti","year":"2020","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2537","DOI":"10.1016\/j.cell.2021.04.015","article-title":"Mechanisms and disease consequences of nonalcoholic fatty liver disease","volume":"184","author":"Loomba","year":"2021","journal-title":"Cell"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1543","DOI":"10.1080\/17512433.2021.1981859","article-title":"Hugan tablets for the treatment of RUCAM based drug-induced liver injury: A propensity score matching analysis using a nationwide database","volume":"14","author":"Li","year":"2021","journal-title":"Expert. Rev. Clin. Pharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1038\/nrgastro.2016.85","article-title":"The role of the gut microbiota in NAFLD","volume":"13","author":"Leung","year":"2016","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"S108","DOI":"10.1093\/cid\/civ177","article-title":"New approaches for bacteriotherapy: Prebiotics, new-generation probiotics, and synbiotics","volume":"60","author":"Patel","year":"2015","journal-title":"Clin. Infect. Dis."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1315","DOI":"10.1080\/17425247.2023.2233900","article-title":"Targeting the gut microbiome to control drug pharmacomicrobiomics: The next frontier in oral drug delivery","volume":"20","author":"Kamath","year":"2023","journal-title":"Expert. Opin. Drug Deliv."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"15718","DOI":"10.1073\/pnas.0407076101","article-title":"The gut microbiota as an environmental factor that regulates fat storage","volume":"101","author":"Ding","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2085","DOI":"10.1136\/gutjnl-2020-323106","article-title":"Effect of green-Mediterranean diet on intrahepatic fat: The DIRECT PLUS randomised controlled trial","volume":"70","author":"Rinott","year":"2021","journal-title":"Gut"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"31259","DOI":"10.1073\/pnas.2017129117","article-title":"Triclosan leads to dysregulation of the metabolic regulator FGF21 exacerbating high fat diet-induced nonalcoholic fatty liver disease","volume":"117","author":"Yueh","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1080\/19490976.2019.1681861","article-title":"Lower gut microbiome diversity and higher abundance of proinflammatory genus Collinsella are associated with biopsy-proven nonalcoholic steatohepatitis","volume":"11","author":"Astbury","year":"2020","journal-title":"Gut Microbes"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1834","DOI":"10.1016\/j.cell.2024.03.014","article-title":"Gut microbiome and metabolome profiling in Framingham heart study reveals cholesterol-metabolizing bacteria","volume":"187","author":"Li","year":"2024","journal-title":"Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1016\/j.cmet.2013.03.013","article-title":"Pleiotropic roles of bile acids in metabolism","volume":"17","author":"Tarling","year":"2013","journal-title":"Cell Metab."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1002\/hep.28709","article-title":"Bile acids and nonalcoholic fatty liver disease: Molecular insights and therapeutic perspectives","volume":"65","author":"Arab","year":"2017","journal-title":"Hepatology"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1837","DOI":"10.1136\/gut.2003.037689","article-title":"Bile secretory function in the obese Zucker rat: Evidence of cholestasis and altered canalicular transport function","volume":"53","author":"Pizarro","year":"2004","journal-title":"Gut"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1885","DOI":"10.1038\/s41591-019-0660-7","article-title":"Three-dimensional spatially resolved geometrical and functional models of human liver tissue reveal new aspects of NAFLD progression","volume":"25","author":"Moser","year":"2019","journal-title":"Nat. Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1146\/annurev-pathol-020117-043617","article-title":"Recent Insights into the Pathogenesis of Nonalcoholic Fatty Liver Disease","volume":"13","author":"Arab","year":"2018","journal-title":"Annu. Rev. Pathol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1002\/hep.28965","article-title":"The ascending pathophysiology of cholestatic liver disease","volume":"65","author":"Jansen","year":"2017","journal-title":"Hepatology"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1016\/j.jhep.2020.07.028","article-title":"Obeticholic acid improves hepatic bile acid excretion in patients with primary biliary cholangitis","volume":"74","author":"Frisch","year":"2021","journal-title":"J. Hepatol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1881","DOI":"10.1136\/gutjnl-2017-314307","article-title":"Suppressed hepatic bile acid signalling despite elevated production of primary and secondary bile acids in NAFLD","volume":"67","author":"Jiao","year":"2018","journal-title":"Gut"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2705","DOI":"10.1016\/S0140-6736(18)31785-9","article-title":"Pegbelfermin (BMS-986036), a PEGylated fibroblast growth factor 21 analogue, in patients with non-alcoholic steatohepatitis: A randomised, double-blind, placebo-controlled, phase 2a trial","volume":"392","author":"Sanyal","year":"2019","journal-title":"Lancet"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"100392","DOI":"10.1016\/j.jhepr.2021.100392","article-title":"Pegbelfermin selectively reduces secondary bile acid concentrations in patients with non-alcoholic steatohepatitis","volume":"4","author":"Luo","year":"2022","journal-title":"JHEP Rep."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2172671","DOI":"10.1080\/19490976.2023.2172671","article-title":"Bile acids as modulators of gut microbiota composition and function","volume":"15","author":"Larabi","year":"2023","journal-title":"Gut Microbes"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"2173","DOI":"10.1128\/jb.173.7.2173-2179.1991","article-title":"Cloning and sequencing of the 7 alpha-hydroxysteroid dehydrogenase gene from Escherichia coli HB101 and characterization of the expressed enzyme","volume":"173","author":"Yoshimoto","year":"1991","journal-title":"J. Bacteriol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"598","DOI":"10.1016\/j.procbio.2014.12.026","article-title":"Two-step enzymatic synthesis of ursodeoxycholic acid with a new 7\u03b2-hydroxysteroid dehydrogenase from Ruminococcus torques","volume":"50","author":"Zheng","year":"2015","journal-title":"Process Biochem."}],"container-title":["Metabolites"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-1989\/15\/7\/433\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:57:54Z","timestamp":1760032674000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-1989\/15\/7\/433"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,24]]},"references-count":51,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["metabo15070433"],"URL":"https:\/\/doi.org\/10.3390\/metabo15070433","relation":{},"ISSN":["2218-1989"],"issn-type":[{"value":"2218-1989","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,24]]}}}